Document Detail

Sphingolipid requirement for generation of a functional v1 component of the vacuolar ATPase.
MedLine Citation:
PMID:  12746460     Owner:  NLM     Status:  MEDLINE    
There has been no previous indication that vacuolar ATPases (V-ATPases) require sphingolipids for function. Here we show, by using Saccharomyces cerevisiae sur4Delta and fen1Delta cells, that sphingolipids with a C26 acyl group are required for generating V1 domains with ATPase activity. Sphingolipids in sur4Delta cells contain C22 and C24 acyl groups instead of C26 acyl groups whereas about 30% of the sphingolipids in fen1Delta cells have C26 acyl groups and the rest have C22 and C24 acyl groups. sur4Delta cells have several phenotypes (vacuolar membrane ATPase, Vma-) that indicate a defect in the V-ATPase, and vacuoles purified from sur4Delta cells have little to no ATPase activity. These phenotypes are less pronounced in fen1Delta cells, consistent with the idea that the C26 acyl group in sphingolipids is necessary for V-ATPase activity. Other results show that the two V-ATPase domains, V1 and V0, are assembled and delivered to the vacuolar membrane in sur4Delta cells similar to wild-type cells. In vitro assembly studies show that V1 from sur4Delta cells associates with wild-type V0 but the complex lacks V-ATPase activity, indicating that V1 is defective. Reciprocal experiments with V0 from sur4Delta cells show that it is normal. We conclude that sphingolipids with a C26 acyl group are required for generating fully functional V1 domains.
Ji-Hyun Chung; Robert L Lester; Robert C Dickson
Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.     Date:  2003-05-13
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  278     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2003 Aug 
Date Detail:
Created Date:  2003-07-28     Completed Date:  2003-09-10     Revised Date:  2011-02-10    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  28872-81     Citation Subset:  IM    
Department of Molecular and Cellular Biochemistry and the Lucille P. Markey Cancer Center, University of Kentucky College of Medicine, Lexington, Kentucky 40536, USA.
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MeSH Terms
Calcium-Transporting ATPases / metabolism
Chromatography, High Pressure Liquid
Electrophoresis, Polyacrylamide Gel
Gene Deletion
Hydrogen-Ion Concentration
Membrane Proteins / genetics
Microscopy, Fluorescence
Protein Subunits / metabolism
Quinacrine / metabolism
Saccharomyces cerevisiae / enzymology*,  genetics
Saccharomyces cerevisiae Proteins / genetics
Spectrometry, Fluorescence
Sphingolipids / chemistry,  genetics,  physiology*
Structure-Activity Relationship
Vacuolar Proton-Translocating ATPases / chemistry*,  genetics,  metabolism*
Grant Support
Reg. No./Substance:
0/Membrane Proteins; 0/Protein Subunits; 0/Saccharomyces cerevisiae Proteins; 0/Sphingolipids; 83-89-6/Quinacrine; EC 2.3.1.-/Acetyltransferases; EC 2.3.1.-/FEN1 protein, S cerevisiae; EC 2.3.1.-/SUR4 protein, S cerevisiae; EC 3.6.1.-/Vacuolar Proton-Translocating ATPases; EC ATPases

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